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A specific qualitative and real-time PCR detection of MON863 maize based on the 5 '-transgene integration sequence

文献类型： 外文期刊

作者： Zhu, Hong 1 ; Zhao, Xiao 2 ; Jia, Junwei 1 ; Sun, Jianping 1 ; Zhao, Kai 1 ;

作者机构： 1.Shanghai Acad Agr Sci, Biotech Inst, Key Lab Agr Genet & Breeding, Shanghai 201106, Peoples R China

2.Xuzhou Normal Univ, Sch Life Sci, Dept Biotech, Xuzhou 221116, Jiangsu Prov, Peoples R China

关键词： LM PCR;Qualitative PCR;TaqMan real-time PCR;MON863

期刊名称：JOURNAL OF CEREAL SCIENCE （ 影响因子：3.616； 五年影响因子：3.891 ）

ISSN： 0733-5210

年卷期： 2008 年 48 卷 3 期

页码：

收录情况： SCI

摘要： Genetically modified crops are widely grown in the world today. Labeling is required when genetically modified organisms (GMOs) are placed on the market. There is a need to establish a specific method for the detection of genetically modified foods. MON863 transgenic maize containing a Cry3Bb1 sequence that produces insecticidal protein cry3Bb1 is a major GMO crop. In this paper, we report studies that designed specific PCR primers and TaqMan probes based upon the 5'-transgene integration sequence, and developed qualitative and quantitative PCR conditions using these primers and probes. We determined the 5'-transgene integration sequence using a ligation-mediated polymerase chain reaction (LM PCR) method. In qualitative PCR studies, the limit of detection (LOD) was 0.5% for MON863 in 100 ng genomic DNA. In the quantitative PCR assays, the limit of detection (LOD) and limit of quantitation (LOQ) are 10 and 100 haploid copies, respectively. Maize samples with different contents of genetically modified component were tested using the established TaqMan real-time PCR system. (C) 2008 Elsevier Ltd. All rights reserved.

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[1]Validation of a rice specific gene, sucrose phosphate synthase, used as the endogenous reference gene for qualitative and real-time quantitative PCR detection of transgenes. Ding, JY,Jia, JW,Yang, LT,Wen, HB,Zhang, CM,Liu, WX,Zhang, DB. 2004

[2]Qualitative and quantitative PCR methods for event-specific detection of genetically modified cotton Mon1445 and Mon531. Yang, LT,Pan, AH,Zhang, KW,Yin, CS,Qian, BJ,Chen, JX,Huang, C,Zhang, DB. 2005

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